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Applications

Application Notes

Validated Applications:WB: 2 - 4 µg/ml dilution

Individual optimization may be required.

For optimal results, primary antibody incubations should be performed at room temperature. The addition of 0.1% Tween 20 to all Blotto solutions may also reduce background. Individual optimization may be required.

Published Applications:ChIPIHC (FFPE)

See references for more information.

Immunogen

This DNMT3A antibody was raised against a full-length protein corresponding to mouse DNMT3A.

Buffer

PBS containing 0.05% sodium azide. Sodium azide is highly toxic.

References

DNMT3A mAb tested by Western blot.Detection of DNMT3A by Western blot. The analysis was performed using DNMT3A mAb at a 2 µg/ml dilution and whole-cell extracts from 293 cells that had been transfected with plasmid expressing either mouse DNMT3A (lane A) or DNMT3B (lane B).

Background

DNMT3A – DNA methyltransferase proteins such as DNMT3A (or DNMT 3A) are involved in DNA methylation, in which a methyl group is added to a cytosine residue on DNA, commonly at the C5 position of a CpG dinucleotide. DNMT3A has been shown to be important in the regulation of specific patterns of DNA methylation. Methylation of mammalian DNA has long been recognized to play a major role in a number of cellular functions such as embryonic development, genetic imprinting, X chromosome inactivation and control of gene expression. DNA methylation is generally associated with transcriptional repression.

Three families of DNMTs have been identified: DNMT1, DNMT2 and DNMT3. The DNMT3 family, containing two active methyltransferases, DNMT3A & DNMT3B, and one DNMT3-Like protein (DNMT3L), establishes the initial CpG methylation pattern de novo and shows the same propensity for methylating unmethylated duplex DNA as for hemi-methylated DNA. DNMT3A and DNMT3B are encoded by two distinct genes and are strongly expressed in embryonic stem cells. DNMT3A is ubiquitous in mammalian cells.

Positive Control

Whole-cell extracts from 293 cells transfected with DNMT3A can be used as a positive control for DNMT3A antibody.

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with DNMT3A antibody (mAb).

Bridging Antibody to Improve ChIP, MeDIP and IP

Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP, MeDIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP, MeDIP and IP experiments that use DNMT3A antibody (mAb).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.